摘要
背景:目前组织工程骨构建研究中的种子细胞主要来源于骨、骨膜、骨髓及骨外组织,近年来的研究多集中于骨髓基质细胞。而脂肪中基质细胞的发现,有望取代骨髓基质细胞。目的:观察体外培养脂肪基质细胞与骨髓基质细胞的生物学特性,并比较二者成骨诱导后的碱性磷酸酶活性,从成骨活性方面来评价脂肪基质细胞能否取代骨髓基质细胞。方法:手术中收集同一人体的脂肪组织与骨髓组织。脂肪组织经机械切割后以Ⅰ型胶原酶消化获得脂肪基质细胞,骨髓组织以淋巴细胞分离液密度梯度离心法分离骨髓基质细胞;体外培养、传代后以诱导培养液行成骨诱导培养。诱导后第2,3周各检测1次细胞中的碱性磷酸酶活性,并行VonKossa钙结节染色鉴定成骨细胞。结果与结论:共获得15例患者的脂肪与骨髓组织,其中10例完成实验。与骨髓基质细胞相比,脂肪基质细胞更易培养成活,扩增速度快;二者细胞形态相似,诱导培养后细胞外基质中均有黑色的钙结节形成;碱性磷酸酶活性二者差异无显著性意义(P>0.05)。结果提示脂肪组织来源丰富,脂肪基质细胞成活容易,具有与骨髓基质细胞相似的生物学性能,且易培养、增殖快,二者的成骨活性相似,脂肪基质细胞比骨髓基质细胞更具有优势。
BACKGROUND：In present research of tissue engineered bone construction,seed cells are mainly harvested from bone,periosteum,bone marrow and non-osseous tissue.Present studies mainly focused on bone marrow stromal cells （BMSCs）,but the discovery of adipose-derived stromal cells （ADSCs） can replace bone marrow stromal cells.OBJECTIVE：To study the biological characteristic of ADSCs and BMSCs cultured in vitro and to compare alkaline phosphatase activities following osteogenesis so as to evaluate whether ADSCs could be a better seed cells to replace BMSCs.METHODS：Adipose tissue and bone marrow from the same body were collected during operation.Adipose tissues were digested with collagenase type Ⅰ to gain ADSCs,and BMSCs were isolated by centrifugation on lymphocyte separation medium.Following in vitro culture and passage,cells were induced to differentiate into osteoblasts.At the second and third weeks after induction,alkaline phosphatase activity in cells was measured for comparison and Von Kossa calcium nodus staining was used to detect osteoblasts.RESULTS AND CONCLUSION：Adipose tissue and bone marrow were gained from 15 patients and 10 of them brought off a successful experiment.Compared with BMSCs,ADSCs can be cultured more easily and proliferate faster.Their morphological characteristics were similar.Black calcium noduses were seen in extracellular matrix.No significant difference was determined in alkaline phosphatase activity between ADSCs and BMSCs （P0.05）.The results proved that adipose tissue is abundant.ADSCs can be cultured more easily,and have similar biological characteristics with BMSCs,and they can be easily cultured,proliferate faster,and can be induced to osteoblasts.ADSCs should be a more ideal seed cells than BMSCs.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第23期4203-4206,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
